LT2 Rule
Cryp tosporidium
         &
      E. coll
  Sample Collection
  Recommendations
  Pocket Crui
   (Printer Friendly Version)

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Cryptosporidium
Preparation for Sample  Filtration

Thoroughly clean* all reused collection equipment and containers (new tubing,
fittings, etc. should not need cleaning), and wear gloves whenever needed to
prevent sample contamination throughout the collection and filtration procedure.
Before connecting the sampling unit to the tap, thoroughly flush stagnant water
and debris in the sample line until the temperature and turbidity stabilize
(approx. 2-3 mins).

Complete the sample collection form including utility and
sample ID information, date, turbidity, pH and temperature.

Pressurized source

1. Determine sample line water pressure at the location described to the State
2. Connect assembled sampling unit, without capsule/foam filter, to sample tap
3. Flush the sampling unit for 2-3 minutes and test for leaks
Unpressurized source

1. Fill 30+ L carboy from a pressurized sample tap or manually collect the sample
  using hose, pump, funnel, etc. from the location described to the State
2. Chill the full carboy to between 1° and 10°C if filtration is not performed
  immediately after filling the carboy
3. Assemble sampling unit, without capsule/foam filter, and insert influent tube
  into the center of the carboy
4. Turn on pump to flush the sampling unit for 2-3 minutes and test for leaks
* Refer to the cleaning procedure located on the Packing and Shipping panel
 of this pocket guide

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Sample Collection Recommendafu
  Cryptosporidium
  Pressurized Sample Filtration
  Enviroehek™ or Envirochek™ HV Capsules
  1. Install filter capsule
  2, Record start time and initial meter reading
  3. Slowly open sample tap, bleed out air, and adjust up to
    the following maximum values:
       • 100 psig and 2 L/min
  4. When at least 10 L has passed through the filter, turn off sample tap
    allowing pressure to decrease until water stops
  5. Record stop time and final meter reading
  6. Hold inlet pointing up, remove outlet tubing and allow water to drain
  7. Open bleed valve to speed the draining process and disconnect inlet tubing
  8. Seal the capsule with vinyl end caps, close the bleed valve, and place in
    gallon-size zippered plastic bag
  9, Chill the sample as soon as possible to between 1° and 10°C
    until ready for shipment

                 Tap pressure >1QO psig
            Pressure
            regulator
Envirochek™   Flow totalizer T
 capsule
                 Tap pressure 1-100 psig

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Filta-Max© Filter
1 . Install foam filter inside filter housing
2. Record start time and initial meter reading
3. Slowly open sample tap and adjust up to the following recommended values:
        • 72.5 psig and 3-4 Umin
4. When at least 1 0 L has passed through the filter, turn off sample tap allowing
  pressure to decrease until water stops
5. Record stop time and final meter reading
6. Hold  inlet pointing up, remove the outlet tubing and allow water to drain
7. Disconnect the intet tubing
8 Seal filter inside filter housing with rubber stoppers and  place into a
  gallon-size zippered plastic bag
9, Chill the sample as soon as possible to between 1° and 10°C
  until ready for shipment
                      Tap pressure >72,5 psig
               Pressure
               regulator
 Filla-Ma*®
filter housing
                    Tap pressure 7.5 - 72,5 psig
                Fife-Max® filter housing

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 Cryptosporidium
 Unpressurized Sample  Filtration
             Envirochek™ or Envirochek™ HV Capsules
1. Install filter capsule and turn on pump
2, Record start time and initial meter reading
3. Bleed out air and adjust flow rate up to a maximum of 2 L/min
4. When at least 10 L has passed through the filter, turn off pump
5. Record stop time and final meter reading
6. Hold inlet pointing up, remove outlet tubing and allow water to drain
7. Open bleed valve to speed the draining process and disconnect inlet tubing
8. Seal the capsule with vinyl end caps, close the bleed valve, and place in
  gallon-size zippered plastic bag
9, Chill the sample as soon as possible to between 1° and 10°C
  until ready for shipment
                       Filta-Max® Filter
               PerisEallic pump
                    I
                                Flow rate meter
                                 with valve x i
1. Install foam filter inside filter housing and
  turn on pump
2. Record start time and initial meter reading
3. Adjust flow rate up to a maximum of 3-4 L/min
4. When at least 10  L has passed through the filter, turn off pump
5. Record stop time  and final meter reading
6. Hold inlet pointing up, remove outlet tubing and allow water to drain
7. Disconnect inlet tubing
8. Seal filter inside filter housing with rubber stoppers and place into a
  gallon-size zippered plastic bag
9. Chili the sample as soon as possible to between 1° and 10°C until ready
  for shipment

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  Cryp tosporidium
  10-L Bulk Water Collection
  Thoroughly clean* all reused collection equipment and containers (new
  hoses, cubitainers, etc, should not need cleaning), and wear gloves
  whenever needed to prevent sample contamination throughout the
  collection procedure. Prior to sample collection, thoroughly flush stagnant
  water and debris in the sample line until the temperature and turbidity
  stabilize (approx. 2-3 mins).
  Complete the sample collection form including utility and sample
  ID information, date, time, turbidity, pH, and temperature.
j
1. Fill the 10-L cubitainer from a pressurized
 sample tap or manually collect sample using
  hose, pump, funnel, etc. from the location
   described to the State.
   2. Once the sample is collected,
   immediately place the cap on the
   cubitainer and tighten
  3. Chill the sample as soon as possible to
 between 1" and 10°C  before shipment
   Refer to the cleaning procedure located on the Packing and
   Shipping panel of this pocket guide

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Sample Collection  Recommendatu
 Cryptosporidium
 Matrix Spike Sample Collection

 Send an extra 10 L bulk sample for spiking at the Cryptosporidium laboratory
 concurrent with the 1st and 21st routine monitoring sample collections and
 again with every 20 samples if more than 40 source water samples are
 collected. Coordinate the collection of matrix spike samples with your laboratory.
 Matrix spike samples should be collected from the same location and at the
 same time as the associated routine sample. The matrix spike sample and
 routine sample should be the same volume. Label the cubitainer with the
 following information using a waterproof pen: PWSID, facility name, and sample
 collection date. Chill the sample as soon as possible to between 1° and 10°C
 before shipment.
 10-L Matrix Spike Sample

 Follow instructions for collecting a
 10-L bulk water sample given on
 previous panel.
 Matrix Spike Samples Greater
 than 10-L
 Either ship the entire matrix spike volume
 as a bulk sample to the laboratory to be spiked and analyzed, OR filter all
 but 10 L of the matrix spike sample at your utility and collect the remaining
 10 L as a bulk sample. Chill both the filter and bulk sample as soon as possible
 to between 1° and 10°C before shipment. Clearly label both the filter and
 bulk sample as comprising 2 parts of a single sample.

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9ns:
ocket Guide
      E.  coli
      Sample Collection

      Wear gloves whenever needed to prevent
      sample contamination throughout the
      collection procedure.

      Complete the sample collection form
      including utility and sample ID
      information, date and time, etc.

      1. Water taps used for sampling
        should be free of aerators, hose
        attachments, etc.
      2. Prior to sample collection, thoroughly
        flush stagnant water and debris
        in the sample line until the
        temperature and turbidity
        stabilize (approx, 2-3 mins):
        DO NOT rinse the sample
        bottle
      3. Record the turbidity,
        pH, and temperature
      4. Aseptically fill the sterile
        E. coli sample bottle
        from a pressurized
        sample tap or manually
        collect sample using,
        hose, pump, funnel,
        etc, from the location
        described to the State
      5. Leave at least one inch of
        head space in the E. coli sample
        bottle, if possible. Collect at least 100 ml of sample for analysis
      6. Immediately close the sample bottle
      7. Chill sample as soon as possible to between 0" and 10'C until ready
        for immediate shipment

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Packing and Shipping
1. Remove sample (filter, cubitainer, or sample bottle) from refrigerator or
  cooler just prior to shipping. Wrap sample in bubble wrap to prevent freezing
2. Place sample in cooler lined with two large plastic trash bags
3. Pack cooler with sealed ice bags or the equivalent number of gel ice packs
  sufficient to maintain the sample between 1° and 10°C during shipment. Use
  the amount of coolant appropriate for the season, geographic location, and
  cooler-type used (follow laboratory recommendations). NEVER USE DRY ICE.
4. Knot and seal the two large plastic trash bags
5. Sign and date sample collection form, place in a water-tight bag in cooler
6. Seal and secure cooler with tape
7. Complete airbill and attach to cooler, retain shipper's copy
8. Ship sample(s) to the laboratory via overnight delivery
    ^Recommended Sampling Equipment Cleaning Procedure
    (or alternatively follow the manufacturer's cleaning instructions)
              Not for sterilization of E. colt sample bottles
  • Thoroughly clean all reused influent equipment prior to the capsule/filter
   housing/bulk water container with a warm detergent solution and scrubbing
   with a brush (new equipment should not need cleaning)
  • Expose equipment to hypochlorite solution (Add 25 mL of 5% household
   bleach for every 1 gallon of pH 7 water) for at least 30 minutes at
   room temperature
  • Thoroughly rinse equipment with reagent grade water that is free of analytes
   and interfering substances. Dry equipment in an area free of potential
   Cryptosporidium contamination

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This pocket guide Is intended as an outline and supplement to the
Source Water Monitoring Guidance Manual for Public Water Systems
and the on-line Sample Collection Training Module. This guide is
intended to provide useful information and recommendations, but
does not substitute for the LT2 Rule.

The guidance manual is available for download and study at:
http://www.epa,gov/safewater/disinfection/lt2/pdfs/guide_lt2_
swmonitoringguidance.pdf

The on-line training module can be accessed at:
http://www.epa.gov/safewater/lt2/training/modules.html

Copies of this pocket guide can be ordered by using the document
number listed below and contacting the National Service Center
for Environmental Publications at 1-800-490-9198
                          Office of Water (46Q1M)
                          EPA 817-K-06-002
                          April  2006

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